Neuroprotection by Recombinant Thrombomodulin

Yuji Taoka; Kenji Okajima; Mitsuhiro Uchiba; Masayoshi Johno

doi:10.1055/s-0037-1613837

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2000; 83(03): 462-468
DOI: 10.1055/s-0037-1613837

Review Article

Schattauer GmbH

Neuroprotection by Recombinant Thrombomodulin

Yuji Taoka

¹From the Departments of Laboratory Medicine and Dermatology, Kumamoto University School of Medicine, Kumamoto, Japan

,

Kenji Okajima

¹From the Departments of Laboratory Medicine and Dermatology, Kumamoto University School of Medicine, Kumamoto, Japan

,

Mitsuhiro Uchiba

¹From the Departments of Laboratory Medicine and Dermatology, Kumamoto University School of Medicine, Kumamoto, Japan

,

Masayoshi Johno

¹From the Departments of Laboratory Medicine and Dermatology, Kumamoto University School of Medicine, Kumamoto, Japan

› Author Affiliations

Abstract

Summary

We examined whether recombinant human soluble thrombomodulin (rhs-TM) reduces compression trauma-induced spinal cord injury through protein C activation in rats. Administration of rhs-TM, either before or after the induction of spinal cord injury (SCI), markedly reduced the resulting motor disturbances. However, neither rhs-TM pretreated with an anti-rhs-TM monoclonal antibody (MAb) F2H5, which inhibits thrombin binding to rhs-TM, nor those pretreated with MAb R5G12, which selectively inhibits protein C activation by rhsTM, prevented the motor disturbances. Intramedullary hemorrhages, observed 24 h after trauma, were significantly reduced in animals given rhs-TM. The increase in the tissue levels of tumor necrosis factor-α (TNF-α), TNF-α mRNA expression, and the accumulation of leukocytes in the damaged segment of the spinal cord were significantly inhibited in animals receiving rhs-TM, but these effects were not observed following administration of rhs-TM pretreated with MAb R5G12 or MAb F2H5. Leukocytopenia and activated protein C all produced effects similar to those of rhs-TM.

These findings suggest that rhs-TM prevents compression traumainduced SCI by inhibiting leukocyte accumulation by reducing the expression of TNF-α mRNA and such therapeutic effects of rhs-TM could be dependent on its protein C activation capacity. Findings further suggest that thrombomodulin can be implicated not only in the coagulation system but in regulation of the inflammatory response.

Keywords

Thrombomodulin - spinal cord injury - activated protein C - tumor necrosis factor-α - leukocytes

Full Text

References

Refrences
1 Owen WG, Esmon CT. Functional properties of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. J Biol Chem 1989; 256: 5532-5.
2 Esmon NL, Owen WG, Esmon CT. Isolation of a membrane-bound cofactor for thrombin-catalyzed activation of protein C. J Biol Chem 1982; 257: 859-64.
3 Esmon CT. The protein C anticoagulant pathway. Arterioscler Thromb 1992; 12: 135-45.
4 Fouw NJ, Jong YE, Haverkate F, Bertina RN. Activated protein C increase fibrin clot lysis by neutralization of plasminogen activator inhibitor; no evidence for a cofactor role of protein S. Thromb Haemost 1988; 60: 328-33.
5 Esmon CT, Taylor Jr FB, Snow TR. Inflammation and coagulation: linked processes potentially regulated through a common pathway mediated by protein C. Thromb Haemost 1991; 66: 160-5.
6 Murakami K, Okajima K, Uchiba M, Johno M, Nakagaki T, Okabe H, Takatsuki K. Activated protein C attenuates endotoxin-induced pulmonary vascular injury by inhibiting activated leukocytes in rats. Blood 1996; 87: 642-7.
7 Murakami K, Okajima K, Uchiba M, Johno M, Nakagaki T, Okabe H, Takatsuki K. Activated protein C prevents LPS-induced pulmonary vascular injury by inhibiting cytokine production. Am J Physiol 1996; 272: L197-2.
8 Uchiba M, Okajima K, Murakami K, Johno M, Okabe H, Takatsuki K. Recombinant thrombomodulin prevents endotoxin-induced lung injury in rats by inhibiting leukocyte activation. Am J Physiol 1996; 271: L470-5.
9 Uchiba M, Okajima K, Murakami K, Johno M, Okabe H, Takatsuki K. rhs-TM prevents ET-induced increase in pulmonary vascular permeability through protein C activation. Am J Physiol 1997; 273: L889-94.
10 Stover SL, Fine PR. The epidemiology and economics of spinal cord injury. Paraplegia 1987; 24: 225-8.
11 Bracken MB, Shepard MJ, Collins WF, Holord TR, Young W, Baskin DS, Eisenberg HM, Flamm E, Leo-Summers L, Maroon L, Maeshall LF, Perot PL, Piepmeier J, Sonntag KH, Wagner FC, Wilberger JE, Winn HR. A randomized, controlled trial of methylprednisolone or naloxone in the treatment of acute spinal-cord injury. New Engl J Med 1990; 322: 1405-11.
12 Young W. Secondary CNS injury. J Neurotrauma 1988; 05: 219-21.
13 Tator CH, Fehlings G. Review of the secondary injury theory of acute spinal cord trauma with emphasis on vascular mechanisms. J Neurosurg 1991; 75: 15-26.
14 Demopoulos HB, Yoder M, Gutman EG, Seligman ML, Flamm ES, Ransohoff J. The fine structure of endothelial surfaces in the microcirculation of experimentally injured feline spinal cords. Scan Electron Microsc 1978; 02: 677-80.
15 Taoka Y, Okajima K, Uchiba M, Murakami K, Kushimoto S, Johno M, Naruo M, Okabe H, Takatsuki K. Role of neutrophils in spinal cord injury in the rat. Neuroscience 1997; 79: 1177-82.
16 Taoka Y, Okajima K, Uchiba M, Murakami K, Harada N, Johno M, Naruo M. Activated protein C reduces the severity of compression- induced spinal cord injury in rats by inhibiting activation of leukocytes. J Neurosci 1998; 18: 1392-8.
17 Gomi K, Zushi M, Honda G, Kawahara S, Matszaki O, Kanabayashi T, Yamamoto S, Maruyama I, Suzuki K. Antithrombotic effect of recombinant human soluble thrombomodulin on thrombin-induced thromboembolism in mice. Blood 1990; 75: 1396-9.
18 Kennet RH. Fusion protocols. fusion by centrifugation of cells suspended in polyethylene glycol. In: Monoclonal Antibodies: Principles and Practice. Kennet R, McKean T, Bechtol K. New York: Plenum; 1980: 365-7.
19 Bajaj SP, Rapaport SI, Prodanos CA. A simplified procedure for purification of human prothrombin, factor IX and factor X. Prep Biochem 1981; 11: 397-412.
20 Nesheim ME, Kettner C, Shaw E, Mann KG. Cofactor dependence of factor Xa incorporation into the prothrombinase complex. J Biol Chem 1981; 256: 6537-40.
21 Taoka Y, Okajima K. Spinal cord injury in the rat. Progress in Neurobiology 1998; 56: 341-58.
22 Taoka Y, Okajima K, Murakami K, Johno M, Naruo M. Role of neutrophil elastase in compression-induced spinal cord injury in rats. Brain Research 1998; 799: 264-9.
23 Rivlin A, Tator CH. Objective clinical assessment of motor function after experimental spinal cord injury in the rat. J Neurosurg 1977; 47: 577-81.
24 Kunkel-Bagden E, Bregman BS. Methods to assess the development and recovery of locomotor function after spinal cord injury in rats. Exp Neurol 1993; 119: 153-64.
25 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
26 Liu S, Adcock IM, Old RV, Barnes PJ, Evans TW. Lipopolysaccharide treatment in vivo induces wide-spread tissue expression of inducible nitric oxide synthase mRNA. Biochem Biophys Res Commun 1993; 196: 1208-13.
27 Taoka Y, Okajima K. Role of leukocytes in spinal cord injury in rats. J Neurotrauma. (in press).
28 Müller-Berghaus G, Eckhart T. The role of granulocytes in the activation of intravascular coagulation and the precipitation of soluble fibrin by endotoxin. Blood 1975; 45: 631-41.
29 Gracia JW, Siflinger-Birnboim NA, Biziors R, Del Vecchio PJ, Fenton II JW, Malik AB. Thrombin-induced increase in albumin permeability across the endothelium. J Cell Physiol 1986; 128: 96-104.
30 Minnear FL, Martin D, Hill L, Taylor AE, Malik AB. Lung morphological and permeability changes induced by intravascular coagulation in dogs. Am J Phyiol 1987; 253: H634-44.
31 Cooper JA, Solano SJ, Bizios R, Kaplan JE, Malik AB. Pulmonary neutrophil kinetics after thrombin-induced intravascular coagulation. J Appl Physiol 1984; 57: 826-32.
32 Bajzar L, Moser J, Nesheim M. TAFI, or plasma procarboxypeptidase B, couples the coagulation and fibrinolytic cascades through the thrombinthrombomodulin complex. J Biol Chem 1996; 271: 16603-8.
33 Yakovlev AG, Faden AI. Sequential expression of c-fos protooncogene, TNF-alpha, and dynorphin genes in spinal cord following experimental traumatic injury. Mol Chem Neuropathol 1994; 23: 179-90.
34 Bartholdi D, Schwab ME. Expression of pro-inflammatory cytokine and chemokine mRNA upon experimental spinal cord injury in mouse: an in situ hybridization study. Eur J Neurosci 1997; 09: 1422-38.
35 Klebanoff SJ, Vadas MA, Harlan JM. Stimulation of neutrophils by tumor necrosis factor. J Immunol 1986; 136: 4220-5.
36 Mulligan MS, Varani J, Dame MK. Role of endothelial-leukocyte adhesion mokecule 1 (ELAM-1) in neutrophil-mediated lung injury in rats. J Clin Invest 1991; 88: 1396-406.
37 Iwasa K, Ikata T, Fukuzawa K. Protective effect of vitamin E on spinal cord injury by compression and concurrent lipid peroxidation. Free Radical Biol Med 1989; 06: 599-606.
38 Nyström B, Berglund JE. Spinal cord restitution following compression injuries in rats. Acta Neurol Scand 1988; 78: 467-72.
39 Schreck R, Albermann K, Baeuerle PA. Nuclear factor kappa B: an oxidative strtess-responsive transcription of eukaryotic cells (a review). Free Radic Res Comunn 1992; 12: 221-37.